姬姆萨色素(又称吉姆萨色素)是由天青II与伊红混合而成，Giemsa染色原理和结果与瑞氏染色基本相同，姬姆萨染色液对胞浆着色力较强，能较好的显示胞浆的嗜碱性程度，特别是对血液和骨髓细胞中的嗜天青、嗜酸性、嗜碱性颗粒，着色清晰，但是对胞核着色偏深,核结构显色不佳，故姬姆萨染液常与瑞氏染液联合使用。
阿拉丁Giemsa Staining Solution以进口的姬姆萨色素、甲醇为主要原料，含特有衬染剂，经研磨配制而成，能呈现出清晰的细胞染色效果，常用于组织切片、血液和细胞涂片、细菌、染色体显带、原生动物寄生虫等染色。嗜酸性颗粒为碱性蛋白质，与酸性染料伊红结合，染粉红色，称为嗜酸性物质；细胞核蛋白和淋巴细胞胞浆为酸性，与碱性染料美蓝或天青结合，染紫蓝色，称为嗜碱性物质；中性颗粒呈等电状态与伊红和美蓝均可结合，染淡紫色，称为中性物质。该试剂仅用于科研领域，不适用于临床诊断或其他用途。
自备材料：
甲醇、乙醇、蒸馏水、0.1%～0.5%乙酸(可选)、显微镜
操作步骤(仅供参考)：
(一)涂片染色
1、常规方法制备血液涂片或骨髓涂片，待涂片自然干燥后，用甲醇固定2～3min。
2、将血液涂片或骨髓涂片放置染色架上，滴加 Giemsa Staining Solution (Ready-to-use) 覆盖涂片，室温或加热染色10～15min。用自来水或蒸馏水从玻片一端缓慢的轻轻冲洗。
3、(可选)0.1%乙酸分化数秒。干燥，镜检。
染色结果：
(二)组织切片染色
1、常规脱水包埋。切片5μm，常规二甲苯或Leagene脱蜡透明液脱蜡至水。
2、蒸馏水清洗2次，每次1～2min。入GiemsaStain浸染12～24h，蒸馏水稍清洗。
3、0.5%乙酸清洗1～2min，自来水稍微冲洗。
4、无水乙醇迅速脱水3次，每次5～10s。
5、二甲苯或Leagene脱蜡透明液透明，中性树脂封固。
染色结果：
注意事项：
1、血液涂片或骨髓涂片应厚薄均匀，否则影响染色效果。
2、涂片染色中Giemsa染色后，请勿先去除染液或直接对涂片用力冲洗。
3、如果染色过深或过浅，应调整染色时间或染色液的浓度。
4、Giemsa涂片染色和组织切片染色中，pH值对染色有一定影响，载玻片应清洁、无酸碱污染，否则影响染色效果。
5、染色液经稀释后液面有金属光泽则表示染液有染色作用，否则染色液可能失效。
6、Giemsa组织切片染色中，染色后需用大量0.1～0.5%乙酸急速冲洗，避免浮面沉淀物污染切片后难以洗脱。
7、0.5%乙酸分化常用于Giemsa组织切片染色，如有必要亦可用于细胞涂片，但其浓度应适量下调，0.5%乙酸分化切片时，切片呈粉红色即可终止。
8、Giemsa组织切片染色中，无水乙醇脱水要迅速，否则切片易褪色。
9、染色液可重复使用，但不宜重复多次，若有沉淀物应过滤后使用。
10、为了您的安全和健康，请穿实验服并戴一次性手套操作。
Giemsa stain (also known as Gimsa stain) is a mixture of Azure II and eosin. The staining principle and results of Giemsa staining are basically consistent with those of Wright's staining. Giemsa staining solution has strong staining affinity for cytoplasm, and can well demonstrate the degree of cytoplasmic basophilia. In particular, it stains the azurophilic, eosinophilic and basophilic granules in blood and bone marrow cells clearly. However, it causes over-staining of cell nuclei and poor visualization of nuclear structures. Therefore, Giemsa staining solution is often used in combination with Wright's staining solution.
Aladdin Giemsa Staining Solution is prepared by grinding, with imported Giemsa stain and methanol as the main raw materials, plus a proprietary mordant. It can produce clear cell staining results, and is commonly used for staining tissue sections, blood and cell smears, bacteria, chromosome banding, protozoan parasites, etc. Eosinophilic granules are alkaline proteins; they bind to the acidic dye eosin and stain pink, which are thus called eosinophilic substances. Nuclear proteins and lymphocyte cytoplasm are acidic; they bind to the basic dyes methylene blue or Azure and stain purple-blue, which are called basophilic substances. Neutral granules are in an isoelectric state and can bind to both eosin and methylene blue, staining pale purple, which are called neutral substances. This reagent is for research use only and is not intended for clinical diagnosis or any other purposes.
Self-prepared Materials:
Methanol, ethanol, distilled water, 0.1%–0.5% acetic acid (optional), microscope
Operating Procedures (For Reference Only):
(I) Smear Staining
1. Prepare a blood smear or bone marrow smear using standard procedures. Allow the smear to air-dry naturally, then fix it with methanol for 2–3 minutes.
2. Place the blood smear or bone marrow smear on a staining rack. Add Giemsa Staining Solution (Ready-to-use) until the smear is completely covered. Stain at room temperature or with heating for 10–15 minutes. Rinse the slide gently and slowly from one end with tap water or distilled water.
3. (Optional) Differentiate with 0.1% acetic acid for several seconds. Air-dry the smear and observe under a microscope.
Staining Results:
Eosinophilic granules
| Pink
|
Basophilic granules
| Purple-blue
|
Neutral granules
| Pale purple
|
(II) Tissue Section Staining
1. Perform routine dehydration and embedding. Cut sections at a thickness of 5 μm, then conduct routine dewaxing to water using xylene or Leagene dewaxing and clearing solution.
2. Rinse twice with distilled water, 1–2 minutes each time. Immerse the sections in Giemsa Stain for 12–24 hours, then rinse briefly with distilled water.
3. Rinse with 0.5% acetic acid for 1–2 minutes, followed by a light rinse with tap water.
4. Dehydrate rapidly three times with anhydrous ethanol, 5–10 seconds each time.
5. Clear the sections with xylene or Leagene dewaxing and clearing solution, then mount and seal with neutral balsam.
Staining Results:
Cell nucleus
| Blue to purple
|
Cytoplasmic cells
| Light blue
|
Connective tissue
| Light red
|
Precautions:
1. Blood smears or bone marrow smears shall be of uniform thickness; otherwise, the staining effect will be affected.
2. After Giemsa staining in smear staining, do not remove the staining solution first or rinse the smear vigorously directly.
3. If the staining is too dark or too light, adjust the staining time or the concentration of the staining solution accordingly.
4. In both Giemsa smear staining and tissue section staining, pH value exerts a certain influence on the staining outcome. Glass slides shall be clean and free of acid-alkali contamination; otherwise, the staining effect will be compromised.
5. If the diluted staining solution shows a metallic luster on the surface, it indicates that the solution is effective for staining; otherwise, the staining solution may have expired.
6. In Giemsa tissue section staining, after staining, rinse the sections rapidly with a large volume of 0.1%–0.5% acetic acid to avoid residual surface precipitates contaminating the sections, which are difficult to wash off subsequently.
7. Differentiation with 0.5% acetic acid is commonly used in Giemsa tissue section staining. If necessary, it can also be applied to cell smears, but its concentration should be appropriately reduced. When differentiating sections with 0.5% acetic acid, terminate the process once the sections turn pink.
8. In Giemsa tissue section staining, dehydration with anhydrous ethanol must be performed rapidly; otherwise, the sections are prone to fading.
9. The staining solution can be reused but not for excessive times. If precipitates are present, filter the solution before use.
10.For your safety and health, please wear a lab coat and disposable gloves during the operation.
ZJ26F0232107
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